NR APAC
AU Villmann,C.; Sandmeier,B.; Pischetsrieder,M.; Becker,C.M.
TI Generation of specific antibodies to detect CNS contaminations in food
QU International Conference - Prion diseases: from basic research to intervention concepts - TSE-Forum, 08.10.-10.10.2003, Gasteig, München - Poster session - BR-97
PT Konferenz-Poster
AB Bovine spongiform encephalopathy (BSE) and the new variant of Creutzfeldt-Jakob disease are believed to be epidemiologically transmitted by the enteral uptake of prion-containing tissue of infected animals. An important means of prophylaxis of bovine and other transmissible spongiform encephalopathies (TSE) is, therefore, eliminating potential prion-containing material from the food chain. We developed a two-step immunological protocol to detect CNS tissue, which can be used for routine food control. Proteolipidprotein (PLP) was identified by database search as a reliable marker protein for CNS tissues. The commercially available antibody recognized an epitope situated in the C-terminal portion of the protein CGRGTKF was not able to recognize the conformation of PLP protein present in food after conventional processing. Therefore, the available antibody appeared to be inappropriate for the use intended here, implying that a new high-affinity antibody was required directed against an epitope resistant against food processing. Criteria defined for antigen selection included: i) CNS-specificity, ii) a particular rather than a soluble antigen, iii) species specificity, iv) immunogenicity. Based on flexibility and hydrophilicity of PLP, a 19 aa epitope in the middle of the protein sequence was chosen for the antibody generation. Besides showing the highest potential immunogenicity within the PLP primary sequence, this motif also harbors a cluster of amino acid substitutions around a basic region near position 120 in chick PLP. The chosen epitope of the antibody against the CNS-specific protein PLP was indeed found to be insensitive to standard processing of food.
AD Carmen Villmann, Cord-Michael Becker, Institut für Biochemie, Emil-Fischer-Zentrum, Universität Erlangen-Nürnberg; Barbara Sandmeier, Monika Pischetsrieder, Institut für Pharmazie und Lebensmittelchemie, Emil-Fischer-Zentrum, Universität Erlangen-Nürnberg
SP englisch
PO Deutschland